TY - JOUR
T1 - Capture and release of cells using a temperature-responsive surface that immobilizes an antibody through DNA duplex formation
AU - Kimura, Tsuyoshi
AU - Nakamura, Naoko
AU - Umeda, Kanji
AU - Hashimoto, Yoshihide
AU - Kishida, Akio
N1 - Funding Information:
This work was supported by Strategic Promotion of Innovative Research and Development from Japan Science and Technology Agency [grant number 1J132]; Japan Society for the Promotion of Science [grant number 16H03181].
PY - 2017/8/13
Y1 - 2017/8/13
N2 - We synthesized a temperature-responsive surface that immobilized an antibody via DNA duplex formation for selective capture and release of target cells. Polyethylene films were modified by grafting poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAAm-co-AAc)), which were prepared at various ratios of NIPAAm/AAc. The increased hydrophilicity of P(NIPAAm-co-PAA) film with decreased temperature was confirmed by water contact angle measurement. Single strand DNA (20mer) was chemically immobilized on the surface and then antibody (anti-mouse CD45, mCD45) modified with the complementary single strand DNA was immobilized on the surface through DNA duplex formation. The mCD45 antibody immobilization was confirmed by immunostaining. HeLa cells (mCD45 negative) and mouse bone marrow (BM) cells (mCD45 positive) were adhered on the surfaces at 37 °C. Although HeLa cells were detached by 4 °C incubation, BM cells were still adhered on the surface and then the adhered cells were released by DNase treatment. From these results, it was suggested that cells could be selectively captured and collected by using a film having surface that immobilizes an antibody via DNA duplex formation.
AB - We synthesized a temperature-responsive surface that immobilized an antibody via DNA duplex formation for selective capture and release of target cells. Polyethylene films were modified by grafting poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAAm-co-AAc)), which were prepared at various ratios of NIPAAm/AAc. The increased hydrophilicity of P(NIPAAm-co-PAA) film with decreased temperature was confirmed by water contact angle measurement. Single strand DNA (20mer) was chemically immobilized on the surface and then antibody (anti-mouse CD45, mCD45) modified with the complementary single strand DNA was immobilized on the surface through DNA duplex formation. The mCD45 antibody immobilization was confirmed by immunostaining. HeLa cells (mCD45 negative) and mouse bone marrow (BM) cells (mCD45 positive) were adhered on the surfaces at 37 °C. Although HeLa cells were detached by 4 °C incubation, BM cells were still adhered on the surface and then the adhered cells were released by DNase treatment. From these results, it was suggested that cells could be selectively captured and collected by using a film having surface that immobilizes an antibody via DNA duplex formation.
KW - Selective capture and release of cell
KW - antibody immobilization
KW - polymer graft
KW - surface modification
KW - temperature-response
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U2 - 10.1080/09205063.2017.1309124
DO - 10.1080/09205063.2017.1309124
M3 - Article
C2 - 28322120
AN - SCOPUS:85016096933
SN - 0920-5063
VL - 28
SP - 1172
EP - 1182
JO - Journal of Biomaterials Science, Polymer Edition
JF - Journal of Biomaterials Science, Polymer Edition
IS - 10-12
ER -